A Phenotypic Screen in Zebrafish Identifies a Novel Small-Molecule Inducer of Ectopic Tail Formation Suggestive of Alterations in Non-Canonical Wnt/PCP Signaling
- Authors
- Gebruers, E., Cordero-Maldonado, M.L., Gray, A.I., Clements, C., Harvey, A.L., Edrada-Ebel, R., de Witte, P.A., Crawford, A.D., and Esguerra, C.V.
- ID
- ZDB-PUB-140213-22
- Date
- 2013
- Source
- PLoS One 8(12): e83293 (Journal)
- Registered Authors
- Cordero-Maldonado, Maria Lorena, Esguerra, Camila V.
- Keywords
- none
- MeSH Terms
-
- Animals
- Bone Morphogenetic Proteins/antagonists & inhibitors
- Bone Morphogenetic Proteins/metabolism
- Coumaric Acids/chemistry
- Coumaric Acids/pharmacology*
- Drug Evaluation, Preclinical
- Embryo, Nonmammalian/embryology*
- Jasminum/chemistry*
- Pyrazoles/pharmacology
- Pyrimidines/pharmacology
- Tail/embryology*
- Wnt Signaling Pathway/drug effects*
- Zebrafish/embryology*
- Zebrafish Proteins/antagonists & inhibitors
- Zebrafish Proteins/metabolism
- PubMed
- 24349481 Full text @ PLoS One
Zebrafish have recently emerged as an attractive model for the in vivo bioassay-guided isolation and characterization of pharmacologically active small molecules of natural origin. We carried out a zebrafish-based phenotypic screen of over 3000 plant-derived secondary metabolite extracts with the goal of identifying novel small-molecule modulators of the BMP and Wnt signaling pathways. One of the bioactive plant extracts identified in this screen – Jasminum gilgianum, an Oleaceae species native to Papua New Guinea – induced ectopic tails during zebrafish embryonic development. As ectopic tail formation occurs when BMP or non-canonical Wnt signaling is inhibited during the tail protrusion process, we suspected a constituent of this extract to act as a modulator of these pathways. A bioassay-guided isolation was carried out on the basis of this zebrafish phenotype, identifying para-coumaric acid methyl ester (pCAME) as the active compound. We then performed an in-depth phenotypic analysis of pCAME-treated zebrafish embryos, including a tissue-specific marker analysis of the secondary tails. We found pCAME to synergize with the BMP-inhibitors dorsomorphin and LDN-193189 in inducing ectopic tails, and causing convergence-extension defects in compound-treated embryos. These results indicate that pCAME may interfere with non-canonical Wnt signaling. Inhibition of Jnk, a downstream target of Wnt/PCP signaling (via morpholino antisense knockdown and pharmacological inhibition with the kinase inhibitor SP600125) phenocopied pCAME-treated embryos. However, immunoblotting experiments revealed pCAME to not directly inhibit Jnk-mediated phosphorylation of c-Jun, suggesting additional targets of SP600125, and/or other pathways, as possibly being involved in the ectopic tail formation activity of pCAME. Further investigation of pCAME’s mechanism of action will help determine this compound’s pharmacological utility.